10736228

pubmed:Citation

4

By testing the sensitivity of Escherichia coli OmpF porin to various natural and synthetic polyamines of different lengths, charge and other molecular characteristics, we were able to identify the molecular properties required for compounds to act as inhibitors of OmpF in the nanomolar range. Inhibitors require at least two amine groups to be effective. For diamines, the optimum length of the hydrocarbon spacer was found to be of eight to ten methylene groups. Triamine molecules based on a 12-carbon motif were found to be more effective that spermidine, an eight-carbon trivalent derivative. But differences in inhibition efficiencies were also found for trivalent compounds depending on the relative position of the internal secondary amine group with respect to the terminal groups. Finally, quaternary ammonium derivatives had no effect, suggesting that the nature of the terminal amine is important for the interaction. From these observations, we deduce that inhibition efficiency in the nanomolar range requires a 12-carbon chain triamine with terminal primary amine groups and replacement of the eighth methylene by a secondary amine. The need for this type of molecular architecture suggests that inhibition is governed by interactions between specific amine groups and protein residues, and that this is not simply due to the accumulation of charges into the pore. Together with previous observations from site-directed mutagenesis studies and inspection of the crystal structure of OmpF, these results allowed us to propose three residues (D113, D121 and Y294) as putative sites of interaction between the channel and spermine. Alanine substitution at each of these three residues resulted in a loss of inhibition by spermine, while mutations of only D113 and D121 affected inhibition by spermidine. Based on these observations, we suggest a model for the molecular determinants involved in the porin-polyamine interaction.

eng

IM

MEDLINE

0022-2836

Copyright 2000 Academic Press.

7

NLM

933-45

pubmed-meshheading:10736228-Amino Acid Substitution, pubmed-meshheading:10736228-Bacterial Proteins, pubmed-meshheading:10736228-Binding Sites, pubmed-meshheading:10736228-Crystallography, X-Ray, pubmed-meshheading:10736228-Dose-Response Relationship, Drug, pubmed-meshheading:10736228-Electric Conductivity, pubmed-meshheading:10736228-Escherichia coli, pubmed-meshheading:10736228-Hydrocarbons, pubmed-meshheading:10736228-Kinetics, pubmed-meshheading:10736228-Liposomes, pubmed-meshheading:10736228-Methane, pubmed-meshheading:10736228-Models, Molecular, pubmed-meshheading:10736228-Molecular Weight, pubmed-meshheading:10736228-Mutagenesis, Site-Directed, pubmed-meshheading:10736228-Polyamines, pubmed-meshheading:10736228-Porins, pubmed-meshheading:10736228-Protein Binding, pubmed-meshheading:10736228-Static Electricity, pubmed-meshheading:10736228-Substrate Specificity

Molecular basis for the polyamine-ompF porin interactions: inhibitor and mutant studies.

Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't